Download Product Insert (PDF)

Total Page：16

File Type：pdf, Size：1020Kb

PRODUCT INFORMATION Bafilomycin A1 Item No. 11038 CAS Registry No.: 88899-55-2 Formal Name: (3Z,5E,7R,8S,9S,11E,13E,15S,16R)-8- hydroxy-16-[(1S,2R,3S)-2-hydroxy-1-methyl- 3-[(2R,4R,5S,6R)-tetrahydro-2,4-dihydroxy- 5-methyl-6-(1-methylethyl)2H-pyran-2-yl] O butyl]-3,15-dimethoxy-5,7,9,11-tetramethyl- OH oxacyclohexadeca-3,5,11,13-tetraen-2-one OH Synonym: NSC 381866 H OH O O MF: C35H58O9 HO FW: 622.8 O O Purity: ≥95% UV/Vis.: λmax: 248, 288 nm Supplied as: A crystalline solid Storage: -20°C Stability: ≥2 years Item Origin: Bacterium/Streptomyces sp. Information represents the product specifications. Batch specific analytical results are provided on each certificate of analysis. Laboratory Procedures Bafilomycin A1 is supplied as a crystalline solid. A stock solution may be made by dissolving the bafilomycin A1 in the solvent of choice, which should be purged with an inert gas. Bafilomycin A1 is soluble in organic solvents such as DMSO and methanol. The solubility of bafilomycin A1 in these solvents is approximately 5 mg/ml. Description Bafilomycin A1 is a fungal metabolite that has been found in Streptomyces and has diverse biological 1-5 + activities. It is an inhibitor of vacuolar H -ATPases (V-ATPases; Ki = 0.5 nM in N. crassa vacuolar membranes) and is greater than 1,000-fold selective for V-ATPases over Na+/K+-, Ca2+-, and H+-ATPases.1,4 Bafilomycin A1 (100 nM) inhibits autophagosome maturation and protein degradation in H-4-II-E cells.2 It inhibits chloroquine-induced apoptosis in primary cerebellar granule neurons (CGNs) but 3 not chloroquine-induced inhibition of macroautophagy. Bafilomycin A1 (100 nM) reduces viral yield in the culture supernatant of Vero E6 and Huh7 cells, as well as HEK293T cells expressing human angiotensin-converting enzyme 2 (ACE2), infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).5 It also reduces lung RNA copy numbers and viral pneumonia in ACE2 transgenic mice infected with SARS-CoV-2 when administered at a dose of 0.1 mg/kg. References 1. Bowman, E.J., Siebers, A., and Altendorf, K. Proc. Natl. Acad. Sci. USA 85(21), 7972-7976 (1988). 2. Yamamoto, A., Tagawa, Y., Yoshimori, T., et al. Cell Struct. Funct. 23(1), 33-42 (1998). 3. Shacka, J.J., Klocke, B.J., and Roth, K.A. Autophagy 2(3), 228-230 (2006). 4. Dröse, S., Bindseil, K.U., Bowman, E.J., et al. Biochemistry 32(15), 3902-3906 (1993). 5. Shang, C., Zhuang, X., Zhang, H., et al. Virol. J. 18(1), 46 (2021). WARNING CAYMAN CHEMICAL THIS PRODUCT IS FOR RESEARCH ONLY - NOT FOR HUMAN OR VETERINARY DIAGNOSTIC OR THERAPEUTIC USE. 1180 EAST ELLSWORTH RD SAFETY DATA ANN ARBOR, MI 48108 · USA This material should be considered hazardous until further information becomes available. Do not ingest, inhale, get in eyes, on skin, or on clothing. Wash thoroughly after handling. Before use, the user must review the complete Safety Data Sheet, which has been sent via email to your institution. PHONE: [800] 364-9897 WARRANTY AND LIMITATION OF REMEDY [734] 971-3335 Buyer agrees to purchase the material subject to Cayman’s Terms and Conditions. Complete Terms and Conditions including Warranty and Limitation of Liability information can be found on our website. FAX: [734] 971-3640 [email protected] Copyright Cayman Chemical Company, 07/26/2021 WWW.CAYMANCHEM.COM.

Copy Link

Recommended publications

Small Molecule Modulators Reveal Mechanisms Regulating Cell Death
Investigating neurodegenerative diseases with small molecule modulators Reka Rebecca Letso Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2011 ©2011 Reka Rebecca Letso All Rights Reserved Abstract Investigating neurodegenerative diseases with small molecule modulators Reka Rebecca Letso Elucidation of the mechanisms underlying cell death in neurodegenerative diseases has proven difficult, due to the complex and interconnected architecture of the nervous system as well as the often pleiotropic nature of these diseases. Cell culture models of neurodegenerative diseases, although seldom recapitulating all aspects of the disease phenotype, enable investigation of specific aspects of these disease states. Small molecule screening in these cell culture models is a powerful method for identifying novel small molecule modulators of these disease phenotypes. Mechanistic studies of these modulators can reveal vital insights into the cellular pathways altered in these disease states, identifying new mechanisms leading to cellular dysfunction, as well as novel therapeutic targets to combat these destructive diseases. Small molecule modulators of protein activity have proven invaluable in the study of protein function and regulation. While inhibitors of protein activity are relatively common, small molecules that can increase protein abundance are quite rare. Small molecule protein upregulators with targeted activities would be of great value in the study of the mechanisms underlying many loss of function diseases. We developed a high- throughput screening approach to identify small molecule upregulators of the Survival of Motor Neuron protein (SMN), whose decreased levels cause the neurodegenerative disease Spinal Muscular Atrophy (SMA). We screened 69,189 compounds for SMN upregulators and performed mechanistic studies on the most active compound, a bromobenzophenone analog designated cuspin-1.
[Show full text]
Protein Kinase C Activates an H' (Equivalent) Conductance in The
Proc. Nati. Acad. Sci. USA Vol. 88, pp. 10816-10820, December 1991 Cell Biology Protein kinase C activates an H' (equivalent) conductance in the plasma membrane of human neutrophils (HI channels/pH regulation/leukocytes/H+-ATPase/bafllomycin) ARVIND NANDA AND SERGIO GRINSTEIN* Division of Cell Biology, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Communicated by Edward A. Adelberg, August 12, 1991 ABSTRACT The rate of metabolic acid generation by it can be calculated that pH1 would be expected to drop by neutrophils increases greatly when they are activated. Intra- over 5 units (to pH 1.6) if the metabolically generated H+ cellular acidification is prevented in part by Na+/H' ex- were to remain within the cell. However, cells stimulated in change, but a sizable component ofH+ extrusion persists in the the nominal absence of Na+ and HCO- maintain their pH1 nominal absence of Na' and HCO3 . In this report we deter- above 6.4, suggesting that alternative (Na+- and HCO3- mined the contribution to H+ extrusion of a putative HI independent) H+ extrusion mechanisms must exist in acti- conductive pathway and its mode ofactivation. In unstimulated vated neutrophils. Accordingly, accelerated extracellular cells, H+ conductance was found to be low and unaffected by acidification can be recorded under these conditions. depolarization. An experimental system was designed to min- The existence of H+-conducting channels has been docu- imize the metabolic acid generation and membrane potential mented in snail neurones (5) and axolotl oocytes (6). In these changes associated with neutrophil activation. By using this tissues, the channels are inactive at the resting plasma system, (3-phorbol esters were shown to increase the H+ membrane potential (Em) but open when the membrane (equivalent) permeability of the plasma membrane.
[Show full text]
Autophagy Flux in Critical Illness, a Translational Approach
www.nature.com/scientificreports OPEN Autophagy fux in critical illness, a translational approach Nicolas Tardif 1,2, Franck Polia2, Inga Tjäder1,2, Thomas Gustafsson3 & Olav Rooyackers1,2 Recent clinical trials suggest that early nutritional support might block the induction of autophagy in Received: 17 October 2018 critically ill patients leading to the development of organ failure. However, the regulation of autophagy, Accepted: 7 June 2019 especially by nutrients, in critical illness is largely unclear. The autophagy fux (AF) in relation to critical Published: xx xx xxxx illness and nutrition was investigated by using an in vitro model of human primary myotubes incubated with serum from critically ill patients (ICU). AF was calculated as the diference of p62 expression in the presence and absence of chloroquine (50 µM, 6 h), in primary myotubes incubated for 24 h with serum from healthy volunteers (n = 10) and ICU patients (n = 93). We observed 3 diferent phenotypes in AF, non-altered (ICU non-responder group), increased (ICU inducer group) or blocked (ICU blocker group). This block was not associate with a change in amino acids serum levels and was located at the accumulation of autophagosomes. The increase in the AF was associated with lower serum levels of non-essential amino acids. Thus, early nutrition during critical illness might not block autophagy but could attenuate the benefcial efect of starvation on reactivation of the autophagy process. This could be of clinical importance in the individual patients in whom this process is inhibited by the critical illness insult. Critically ill patients treated in an Intensive Care Unit (ICU) ofen have organ failure already at their admittance or will develop it early during their stay in the unit.
[Show full text]
Gabaergic Signaling Linked to Autophagy Enhances Host Protection Against Intracellular Bacterial Infections
ARTICLE DOI: 10.1038/s41467-018-06487-5 OPEN GABAergic signaling linked to autophagy enhances host protection against intracellular bacterial infections Jin Kyung Kim1,2,3, Yi Sak Kim1,2,3, Hye-Mi Lee1,3, Hyo Sun Jin4, Chiranjivi Neupane 2,5, Sup Kim1,2,3, Sang-Hee Lee6, Jung-Joon Min7, Miwa Sasai8, Jae-Ho Jeong 9,10, Seong-Kyu Choe11, Jin-Man Kim12, Masahiro Yamamoto8, Hyon E. Choy 9,10, Jin Bong Park 2,5 & Eun-Kyeong Jo1,2,3 1234567890():,; Gamma-aminobutyric acid (GABA) is the principal inhibitory neurotransmitter in the brain; however, the roles of GABA in antimicrobial host defenses are largely unknown. Here we demonstrate that GABAergic activation enhances antimicrobial responses against intracel- lular bacterial infection. Intracellular bacterial infection decreases GABA levels in vitro in macrophages and in vivo in sera. Treatment of macrophages with GABA or GABAergic drugs promotes autophagy activation, enhances phagosomal maturation and antimicrobial responses against mycobacterial infection. In macrophages, the GABAergic defense is mediated via macrophage type A GABA receptor (GABAAR), intracellular calcium release, and the GABA type A receptor-associated protein-like 1 (GABARAPL1; an Atg8 homolog). Finally, GABAergic inhibition increases bacterial loads in mice and zebraﬁsh in vivo, sug- gesting that the GABAergic defense plays an essential function in metazoan host defenses. Our study identiﬁed a previously unappreciated role for GABAergic signaling in linking antibacterial autophagy to enhance host innate defense against intracellular bacterial infection. 1 Department of Microbiology, Chungnam National University School of Medicine, Daejeon 35015, Korea. 2 Department of Medical Science, Chungnam National University School of Medicine, Daejeon 35015, Korea.
[Show full text]
Astrocytes: a Driving Force in Brain Signaling and Encephalopathies Aleksandar Jeremic Iowa State University
Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2001 Astrocytes: a driving force in brain signaling and encephalopathies Aleksandar Jeremic Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Neuroscience and Neurobiology Commons, and the Neurosciences Commons Recommended Citation Jeremic, Aleksandar, "Astrocytes: a driving force in brain signaling and encephalopathies " (2001). Retrospective Theses and Dissertations. 648. https://lib.dr.iastate.edu/rtd/648 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps.
[Show full text]
Cisplatin-Induced Apoptosis Inhibits Autophagy, Which Acts As a Pro-Survival Mechanism in Human Melanoma Cells
Cisplatin-Induced Apoptosis Inhibits Autophagy, Which Acts as a Pro-Survival Mechanism in Human Melanoma Cells Barbara Del Bello, Marzia Toscano, Daniele Moretti, Emilia Maellaro* Department of Pathophysiology, Experimental Medicine and Public Health, Istituto Toscano Tumori, University of Siena, Siena, Italy Abstract The interplay between a non-lethal autophagic response and apoptotic cell death is still a matter of debate in cancer cell biology. In the present study performed on human melanoma cells, we investigate the role of basal or stimulated autophagy in cisplatin-induced cytotoxicity, as well as the contribution of cisplatin-induced activation of caspases 3/7 and conventional calpains. The results show that, while down-regulating Beclin-1, Atg14 and LC3-II, cisplatin treatment inhibits the basal autophagic response, impairing a physiological pro-survival response. Consistently, exogenously stimulated autophagy, obtained with trehalose or calpains inhibitors (MDL-28170 and calpeptin), protects from cisplatin-induced apoptosis, and such a protection is reverted by inhibiting autophagy with 3-methyladenine or ATG5 silencing. In addition, during trehalose-stimulated autophagy, the cisplatin-induced activation of calpains is abrogated, suggesting the existence of a feedback loop between the autophagic process and calpains. On the whole, our results demonstrate that in human melanoma cells autophagy may function as a beneficial stress response, hindered by cisplatin-induced death mechanisms. In a therapeutic perspective, these findings suggest that the efficacy of cisplatin-based polychemotherapies for melanoma could be potentiated by inhibitors of autophagy. Citation: Del Bello B, Toscano M, Moretti D, Maellaro E (2013) Cisplatin-Induced Apoptosis Inhibits Autophagy, Which Acts as a Pro-Survival Mechanism in Human Melanoma Cells.
[Show full text]
Na+-Dependent and Na+-Independent Mechanisms for Inorganic Phosphate Uptake in Trypanosoma Rangeli
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1820 (2012) 1001–1008 Contents lists available at SciVerse ScienceDirect Biochimica et Biophysica Acta journal homepage: www.elsevier.com/locate/bbagen Na+-dependent and Na+-independent mechanisms for inorganic phosphate uptake in Trypanosoma rangeli C.F. Dick a,b,c, A.L.A. Dos-Santos b,c, D. Majerowicz d,e, K.C. Gondim d,e, C. Caruso-Neves f, I.V. Silva g, A. Vieyra f,c, J.R. Meyer-Fernandes b,c,⁎ a Instituto de Microbiologia Professor Paulo de Góes, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil b Laboratório de Bioquímica Celular, Instituto de Bioquímica Medica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil c Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagem, Rio de Janeiro, RJ, Brazil d Laboratório de Bioquímica e Fisiologia de Insetos, Instituto de Bioquímica Medica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil e Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, RJ, Brazil f Instituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil g Departamento de Morfologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, ES, Brazil article info abstract Article history: Background: Trypanosoma rangeli is dependent on the presence of exogenous orthophosphate (Pi) for maxi- Received 31 August 2011 mal growth and ecto-phosphatase activity is responsible for Pi supply under low Pi.
[Show full text]
Amino Acid Transporters on the Guard of Cell Genome and Epigenome
cancers Review Amino Acid Transporters on the Guard of Cell Genome and Epigenome U˘gurKahya 1,2 , Ay¸seSedef Köseer 1,2,3,4 and Anna Dubrovska 1,2,3,4,5,* 1 OncoRay–National Center for Radiation Research in Oncology, Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, Helmholtz-Zentrum Dresden-Rossendorf, 01309 Dresden, Germany; [email protected] (U.K.); [email protected] (A.S.K.) 2 Helmholtz-Zentrum Dresden-Rossendorf, Institute of Radiooncology-OncoRay, 01328 Dresden, Germany 3 National Center for Tumor Diseases (NCT), Partner Site Dresden and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany 4 Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany 5 German Cancer Consortium (DKTK), Partner Site Dresden and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany * Correspondence: [email protected]; Tel.: +49-351-458-7150 Simple Summary: Amino acid transporters play a multifaceted role in tumor initiation, progression, and therapy resistance. They are critical to cover the high energetic and biosynthetic needs of fast- growing tumors associated with increased proliferation rates and nutrient-poor environments. Many amino acid transporters are highly expressed in tumors compared to the adjacent normal tissues, and their expression correlates with tumor progression, clinical outcome, and treatment resistance. Tumor growth is driven by epigenetic and metabolic reprogramming and is associated with excessive production of reactive oxygen species causing the damage of vital macromolecules, including DNA. This review describes the role of the amino acid transporters in maintaining tumor redox homeostasis, DNA integrity, and epigenetic landscape under stress conditions and discusses them as potential targets for tumor imaging and treatment.
[Show full text]
Engineering Bafilomycin High-Producers by Manipulating Regulatory and Biosynthetic Genes in the Marine Bacterium Streptomyces Lo
marine drugs Article Engineering Bafilomycin High-Producers by Manipulating Regulatory and Biosynthetic Genes in the Marine Bacterium Streptomyces lohii Zhong Li 1,2,3, Shuai Li 1, Lei Du 1 , Xingwang Zhang 1, Yuanyuan Jiang 1,2,3, Wenhua Liu 1, Wei Zhang 1 and Shengying Li 1,4,* 1 State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China; [email protected] (Z.L.); [email protected] (S.L.); [email protected] (L.D.); [email protected] (X.Z.); [email protected] (Y.J.); [email protected] (W.L.); [email protected] (W.Z.) 2 Shandong Provincial Key Laboratory of Synthetic Biology, CAS Key Laboratory of Biofuels at Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China 3 College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China 4 Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China * Correspondence: [email protected] Abstract: Bafilomycin A1 is the representative compound of the plecomacrolide natural product family. This 16-membered ring plecomacrolide has potent antifungal and vacuolar H+-ATPase inhibitory activities. In our previous work, we identified a bafilomycin biosynthetic gene cluster (baf ) from the marine bacterium Streptomyces lohii ATCC BAA-1276, wherein a luxR family regulatory gene orf1 and an afsR family regulatory gene bafG were revealed based on bioinformatics analysis. In this study, the positive regulatory roles of orf1 and bafG for bafilomycin biosynthesis are characterized through gene inactivation and overexpression. Compared to the wild-type S.
[Show full text]
ATG4D Is the Main ATG8 Delipidating Enzyme in Mammalian Cells and Protects Against Cerebellar Neurodegeneration
Cell Death & Differentiation (2021) 28:2651–2672 https://doi.org/10.1038/s41418-021-00776-1 ARTICLE ATG4D is the main ATG8 delipidating enzyme in mammalian cells and protects against cerebellar neurodegeneration 1,2,3 1,2,3 3 2,3 1,2 Isaac Tamargo-Gómez ● Gemma G. Martínez-García ● María F. Suárez ● Verónica Rey ● Antonio Fueyo ● 1,3 2,3,4 2,3,4,5 6 6 Helena Codina-Martínez ● Gabriel Bretones ● Xurde M. Caravia ● Etienne Morel ● Nicolas Dupont ● 7 1,3 8,9 8,9 6 Roberto Cabo ● Cristina Tomás-Zapico ● Sylvie Souquere ● Gerard Pierron ● Patrice Codogno ● 2,3,4,5 1,2,3 1,2,3 Carlos López-Otín ● Álvaro F. Fernández ● Guillermo Mariño Received: 3 July 2020 / Revised: 16 March 2021 / Accepted: 17 March 2021 / Published online: 1 April 2021 © The Author(s) 2021. This article is published with open access, corrected publication 2021 Abstract Despite the great advances in autophagy research in the last years, the speciﬁc functions of the four mammalian Atg4 proteases (ATG4A-D) remain unclear. In yeast, Atg4 mediates both Atg8 proteolytic activation, and its delipidation. However, it is not clear how these two roles are distributed along the members of the ATG4 family of proteases. We show that these two functions are preferentially carried out by distinct ATG4 proteases, being ATG4D the main delipidating 1234567890();,: 1234567890();,: enzyme. In mammalian cells, ATG4D loss results in accumulation of membrane-bound forms of mATG8s, increased cellular autophagosome number and reduced autophagosome average size. In mice, ATG4D loss leads to cerebellar neurodegeneration and impaired motor coordination caused by alterations in trafﬁcking/clustering of GABAA receptors.
[Show full text]
Pharmacological Modulators of Autophagy Activate a Parallel Noncanonical Pathway Driving Unconventional LC3 Lipidation
AUTOPHAGY 2017, VOL. 13, NO. 5, 854–867 http://dx.doi.org/10.1080/15548627.2017.1287653 BASIC RESEARCH PAPER Pharmacological modulators of autophagy activate a parallel noncanonical pathway driving unconventional LC3 lipidation Elise Jacquina,Stephanie Leclerc-Mercierb, Celine Judonc, Emmanuelle Blanchardd,e, Sylvie Fraitagb, and Oliver Floreya aSignalling Programme, The Babraham Institute, Babraham, UK; bDepartment of Pathology, Necker-Enfants Malades Hospital, MAGEC-Necker Team, Paris, France; cUniversite de Franche-Comte, Besancon, France; dCentre Hospitalier Regional Universitaire, University Francois-Rabelais,¸ Faculty of Medicine, Tours, France; eINSERM, U966, Tours, France ABSTRACT ARTICLE HISTORY The modulation of canonical macroautophagy/autophagy for therapeutic benefit is an emerging strategy Received 1 August 2016 of medical and pharmaceutical interest. Many drugs act to inhibit autophagic flux by targeting lysosome Revised 9 January 2017 function, while others were developed to activate the pathway. Here, we report the surprising finding that Accepted 23 January 2017 many therapeutically relevant autophagy modulators with lysosomotropic and ionophore properties, KEYWORDS classified as inhibitors of canonical autophagy, are also capable of activating a parallel noncanonical autophagy; LAP; lipidation; autophagy pathway that drives MAP1LC3/LC3 lipidation on endolysosomal membranes. Further, we provide lysosome; lysosomotropic; the first evidence supporting drug-induced noncanonical autophagy in vivo using the local anesthetic noncanonical lidocaine and human skin biopsies. In addition, we find that several published inducers of autophagy and mitophagy are also potent activators of noncanonical autophagy. Together, our data raise important issues regarding the interpretation of LC3 lipidation data and the use of autophagy modulators, and highlight the need for a greater understanding of the functional consequences of noncanonical autophagy.
[Show full text]
Characterization of Bafilomycin Biosynthesis In
The Journal of Antibiotics (2017) 70, 616–624 & 2017 Japan Antibiotics Research Association All rights reserved 0021-8820/17 www.nature.com/ja ORIGINAL ARTICLE Characterization of bafilomycin biosynthesis in Kitasatospora setae KM-6054 and comparative analysis of gene clusters in Actinomycetales microorganisms Ayako Nara1,3, Takuya Hashimoto2,3, Mamoru Komatsu1, Makoto Nishiyama2, Tomohisa Kuzuyama2 and Haruo Ikeda1 Bafilomycins A1,C1 and B1 (setamycin) produced by Kitasatospora setae KM-6054 belong to the plecomacrolide family, which exhibit antibacterial, antifungal, antineoplastic and immunosuppressive activities. An analysis of gene clusters from K. setae KM-6054 governing the biosynthesis of bafilomycins revealed that it contains five large open reading frames (ORFs) encoding the multifunctional polypeptides of bafilomycin polyketide synthases (PKSs). These clustered PKS genes, which are responsible for bafilomycin biosynthesis, together encode 11 homologous sets of enzyme activities, each catalyzing a specific round of polyketide chain elongation. The region contains an additional 13 ORFs spanning a distance of 73 287 bp, some of which encode polypeptides governing other key steps in bafilomycin biosynthesis. Five ORFs, BfmB, BfmC, BfmD, BfmE and BfmF, were involved in the formation of methoxymalonyl-acyl carrier protein (ACP). Two possible regulatory genes, bfmR and bfmH, were found downstream of the above genes. A gene-knockout analysis revealed that BfmR was only a transcriptional regulator for the transcription of bafilomycin biosynthetic genes. Two genes, bfmI and bfmJ, were found downstream of bfmH. An analysis of these gene-disruption mutants in addition to an enzymatic analysis of BfmI and BfmJ revealed that BfmJ activated fumarate and BfmI functioned as a catalyst to form a fumaryl ester at the C21 hydroxyl residue of bafilomycin A1.
[Show full text]